Two novel autophosphorylation sites in the juxtamembrane region of the PDGF alpha-receptor, Tyr-572 and Tyr-574, were identified. A Y572/574F mutant PDGF (alpha)-receptor was generated and stably expressed in porcine aortic endothelial cells. In contrast to the wild-type receptor, the mutant receptor was unable to associate with or activate Src family tyrosine kinases. Tyrosine phosphorylated synthetic peptides representing the juxtamembrane sequence of the receptor dose-dependently inhibited the binding of Src family tyrosine kinases to the autophosphorylated PDGF alpha-receptor. The mutant receptor showed similar PDGF-induced kinase activity and ability to mediate mitogenicity, actin reorganization and chemotaxis as the wild-type receptor. Thus activation of Src family kinases by the PDGF alpha-receptor is not essential for PDGF-induced mitogenicity or actin reorganization.

Auerbach
R.
,
Auerbach
W.
,
Polakowski
I.
(
1991
).
Assays for angiogenesis: A review.
Pharmac. Ther
51
,
1
11
Barone
M. V.
,
Courtneidge
S. A.
(
1995
).
Myc but not Fos rescue of PDGF signalling block caused by kinase-inactive Src.
Nature
378
,
509
512
Bazenet
C. E.
,
Gelderloos
J. A.
,
Kazlauskas
A.
(
1996
).
Phosphorylation of tyrosine 720 in the platelet-derived growth factor alpha receptor is required for binding of Grb2 and SHP-2 but not for activation of Ras or cell proliferation.
Mol. Cell Biol
16
,
6926
6936
Boström
H.
,
Willetts
K.
,
Pekny
M.
,
Leveen
P.
,
Lindahl
P.
,
Hedstrand
H.
,
Pekna
M.
,
Hellström
M.
,
Gebre-Medhin
S.
,
Schalling
M.
,
Nilsson
M.
,
Kurland
S.
,
Törnell
J.
,
Heath
J. K.
,
Betsholtz
C.
(
1996
).
PDGF-A signaling is a critical event in lung alveolar myofibroblast development and alveogenesis.
Cell
85
,
863
873
Kapeller
R.
,
Soltoff
S.
(
1991
).
Oncogenes and signal transduction.
Cell
64
,
281
302
Claesson-Welsh
L.
,
Eriksson
A.
,
Moren
A.
,
Severinsson
L.
,
Ek
B.
,
Östman
A.
,
Betsholtz
C.
,
Heldin
C.-H.
(
1988
).
cDNA cloning and expression of a human platelet-derived growth factor (PDGF) receptor specific for B-chain-containing PDGF molecules.
Mol. Cell. Biol
8
,
3476
3486
Claesson-Welsh
L.
,
Eriksson
A.
,
Westermark
B.
,
Heldin
C.-H.
(
1989
).
cDNA cloning and expression of the human A-type platelet-derived growth factor (PDGF) receptor establishes structural similarity to the B-type PDGF receptor.
Proc. Nat. Acad. Sci. USA
86
,
4917
4921
Claesson-Welsh
L.
(
1994
).
Platelet-derived growth factor receptor signals.
J. Biol. Chem
269
,
32023
32026
Egan
S. E.
,
Giddings
B. W.
,
Brooks
M. W.
,
Buday
L.
,
Sizeland
A. M.
,
Weinberg
R. A.
(
1993
).
Association of Sos Ras exchange protein with Grb2 is implicated in tyrosine kinase signal transduction and transformation.
Nature
363
,
45
51
Eisenman
R. N.
,
Cooper
J. A.
(
1995
).
Beating a path to Myc.
Nature
378
,
438
439
Eriksson
A.
,
Nånberg
E.
,
Rönnstrand
L.
,
Engström
U.
,
Hellman
U.
,
Rupp
E.
,
Carpenter
G.
,
Heldin
C.-H.
,
Claesson-Welsh
L.
(
1995
).
Demonstration of functionally different interactions between phospholipase C-and the two types of platelet-derived growth factor receptors.
J. Biol. Chem
270
,
7773
7781
Eriksson
A.
,
Siegbahn
A.
,
Westermark
B.
,
Heldin
C.-H.
,
Claesson-Welsh
L.
(
1992
).
PDGF-and -receptors activate unique and common signal transduction pathways.
EMBO J
11
,
543
550
Heldin
C.-H.
,
Bäckström
G.
,
Östman
A.
,
Hammacher
A.
,
Rönnstrand
L.
,
Rubin
K.
,
Nister
M.
,
Westermark
B.
(
1988
).
Binding of different dimeric forms of PDGF to human fibroblasts: evidence for two separate receptor types.
EMBO J
7
,
1387
1393
Kypta
R. M.
,
Goldberg
Y.
,
Ulug
E. T.
,
Courtneidge
S. A.
(
1990
).
Association between the PDGF receptor and members of the src family of tyrosine kinases.
Cell
62
,
481
492
Leveen
P.
,
Pekny
M.
,
Gebre-Medhin
S.
,
Swolin
B.
,
Larsson
E.
,
Betsholtz
C.
(
1994
).
Mice deficient for PDGF B show renal, cardiovascular and hematological abnormalities.
Genes Dev
8
,
1875
1887
Liao
F.
,
Shin
H. S.
,
Rhee
S. G.
(
1993
).
In vitro tyrosine phosphorylation of PLC-1 and PLC- 2 by src-family protein tyrosine kinases.
Biochem. Biophys. Res. Commun
191
,
1028
1033
Marais
R.
,
Light
Y.
,
Paterson
H. F.
,
Marshall
C. J.
(
1995
).
Ras recruits Raf-1 to the plasma membrane for activation by tyrosine phosphorylation.
EMBO J
14
,
3136
3145
Matsui
T.
,
Heidaran
M.
,
Miki
T.
,
Toru
M.
,
Popescu
N.
,
La Rochelle
W.
,
Kraus
M.
,
Pierce
J.
,
Aaronson
S. A.
(
1989
).
Isolation of a novel receptor cDNA establishes the existence of two PDGF receptor genes.
Science
243
,
800
803
McGlade
J.
,
Cheng
A.
,
Pelicci
G.
,
Pelicci
P. G.
,
Pawson
T.
(
1992
).
Shc proteins are phosphorylated and regulated by the v-Src and v-Fps protein-tyrosine kinases.
Proc. Nat. Acad. Sci. USA
89
,
8869
8873
Miyazono
K.
,
Okabe
T.
,
Urabe
A.
,
Takaku
F.
,
Heldin
C.-H.
(
1987
).
Purification and properties of an endothelial cell growth factor from human platelets.
J. Biol. Chem
262
,
4098
4103
Mori
S.
,
Rönnstrand
L.
,
Yokote
K.
,
Engström
Å.
,
Courtneidge
S. A.
,
Claesson-Welsh
L.
,
Heldin
C.-H.
(
1993
).
Identification of two juxtamembrane autophosphorylation sites in the PDGF-receptor; involvement in the interaction with Src family tyrosine kinases.
EMBO J
12
,
2257
2264
Nada
S.
,
Okada
M.
,
Aizawa
S.
,
Nakagawa
H.
(
1994
).
Identification of major tyrosine-phosphorylated proteins in Csk-deficient cells.
Oncogene
9
,
3571
3578
Nakanishi
O.
,
Shibasaki
F.
,
Hidaka
M.
,
Homma
Y.
,
Takenawa
T.
(
1993
).
Phospholipase C-1 associates with viral and cellular src kinases.
J. Biol. Chem
268
,
10754
10759
Okada
M.
,
Nada
S.
,
Yamanashi
Y.
,
Yamamoto
T.
,
Nakagawa
H.
(
1991
).
CSK: a protein-tyrosine kinase involved in regulation of src family kinases.
J. Biol. Chem
266
,
24249
24252
Pawson
T.
(
1995
).
Protein modules and signalling networks.
Nature
373
,
573
580
Rupp
E.
,
Siegbahn
A.
,
Rönnstrand
L.
,
Wernstedt
C.
,
Claesson-Welsh
L.
,
Heldin
C.-H.
(
1994
).
A unique autophosphorylation site in the platelet-derived growth factorreceptor from a heterodimeric receptor complex.
Eur. J. Biochem
225
,
29
41
Rönnstrand
L.
,
Mori
S.
,
Arvidsson
A.-K.
,
Eriksson
A.
,
Wernstedt
C.
,
Hellman
U.
,
Claesson-Welsh
L.
,
Heldin
C.-H.
(
1992
).
Identification of two C-terminal autophosphorylation sites in the PDGF-receptor: involvement in the interaction with phospholipase C-.
EMBO J
11
,
3911
3919
Smith
M. R.
,
DeGudicubus
S. J.
,
Stacey
D. W.
(
1986
).
Requirement for c-ras proteins during viral oncogene transformation.
Nature
320
,
540
543
Soriano
P.
(
1994
).
Abnormal kidney development and hematological disorders in PDGF-receptor mutant mice.
Genes Dev
8
,
1888
1896
Soriano
P.
(
1997
).
The PDGFreceptor is required for neural crest cell development and for normal patterning of the somites.
Development
124
,
2691
2700
Stokoe
D.
,
McCormick
F.
(
1997
).
Activation of c-Raf-1 by Ras and Src through different mechanisms: activation in vivo and in vitro.
EMBO J
16
,
2384
2396
Twamley-Stein
G. M.
,
Pepperkok
R.
,
Ansorge
W.
,
Courtneidge
S. A.
(
1993
).
The Src family tyrosine kinases are required for platelet-derived growth factor-mediated signal transduction in NIH 3T3 cells.
Proc. Nat. Acad. Sci. USA
90
,
7696
7700
Ullrich
A.
,
Schlessinger
J.
(
1990
).
Signal transduction by receptors with tyrosine kinase activity.
Cell
61
,
203
212
Vaillancourt
R. R.
,
Heasley
L. E.
,
Zamarripa
J.
,
Storey
B.
,
Valius
M.
,
Kazlauskas
A.
,
Johnson
G. L.
(
1995
).
Mitogen-activated protein kinase activation is insufficient for growth factor receptor-mediated PC12 cell differentiation.
Mol. Cell. Biol
15
,
3644
3653
Wennström
S.
,
Landgren
E.
,
Blume-Jensen
P.
,
Claesson-Welsh
L.
(
1992
).
The platelet-derived growth factor-receptor kinase insert confers specific signaling properties to a chimeric fibroblast growth factor receptor.
J. Biol. Chem
267
,
13749
13756
Wennström
S.
,
Siegbahn
A.
,
Yokote
K.
,
Arvidsson
A.-K.
,
Heldin
C.-H.
,
Mori
S.
,
Claesson-Welsh
L.
(
1994
).
Membrane ruffling and chemotaxis transduced by the PDGF-receptor require the binding site for phosphatidylinositol 3 kinase.
Oncogene
9
,
651
660
Yarden
Y.
,
Escobedo
J. A.
,
Kuang
W.-J.
,
Yang-Feng
T. L.
,
Daniel
T. O.
,
Tremble
P. M.
,
Chen
E. Y.
,
Ando
M. E.
,
Harkins
R. N.
,
Francke
U.
,
Friend
V. A.
,
Ullrich
A.
,
Williams
L. T.
(
1986
).
Structure of the receptor for platelet-derived growth factor helps define a family of closely related growth factor receptors.
Nature
323
,
226
232
Yokote
K.
,
Mori
S.
,
Siegbahn
A.
,
Rönnstrand
L.
,
Wernstedt
C.
,
Heldin
C.-H.
,
Claesson-Welsh
L.
(
1996
).
Structural determinants in the platelet-derived growth factor-receptor implicated in modulation of chemotaxis.
J. Biol. Chem
271
,
5101
5111
Yu
J.-C.
,
Heidaran
M. A.
,
Pierce
J. H.
,
Gutkind
J. S.
,
Lombardi
D.
,
Ruggiero
M.
,
Aaronson
S. A.
(
1991
).
Tyrosine mutations within theplatelet-derived growth factor receptor kinase insert domain abrogate receptor-associated phosphatidylinositol-3 kinase activity without affecting mitogenic or chemotactic signal transduction.
Mol. Cell. Biol
11
,
3780
3785
This content is only available via PDF.